Contents

Contributors

Preface

Glossary of Terms

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PART I: IDENTIFICATION AND DIAGNOSTICS

1 Molecular Approaches to Virus Characterization and Detection

1.1. Introduction

1.2. Current Virus Taxonomy and Classification

1.3. Preliminary Approaches to Virus Identification

1.4. Methodology for Virus Isolation and Fractionation (Nucleic Acid and Protein Purification Techniques)

1.5. Biochemical/Molecular Approaches to Virus Identification and Diagnosis

1.6. PCR and Virus-specific Primer Development

1.7. Conclusions

2 Molecular Approaches and Techniques for the Study of Entomopathogenic Bacteria

2.1. Introduction

2.2. Classification of Entomopathogenic Bacteria

2.3. Conclusions

3 Molecular Methods for Identification and Diagnosis of Fungi

3.1. Introduction

3.2. General Considerations

3.3. Genetic Fingerprinting

3.4. DNA Sequencing

3.5. Diagnosis and Detection

3.6. Conclusions and Future Prospects

4 Molecular Approaches and the Taxonomy of Insect-parasitic and Pathogenic Nematodes

4.1. Introduction

4.2. Nematode Diagnosis and the Barcode System

4.3. A Review of Molecular Approaches Considered for Insect-parasitic and Pathogenic Nematode Taxonomy

4.4. Techniques Considered for Obtaining DNA Sequences

4.5. Sequencing

4.6. Sequence Manipulation and Analysis

4.7. Conclusions

5 Identification and Diagnostics of Entomopathogenic Protozoa

5.1. Introduction

5.2. Molecular Identification of Species and Strains

5.3. Conclusion and Future Perspectives

PART II: EVOLUTIONARY RELATIONSHIPS AND POPULATION GENETICS

6 Phylogenetic Studies with Entomopathogenic Bacteria with Special Emphasis on Symbionts of Entomopathogenic Nematodes

6.1. Introduction

6.2. Genes Considered

6.3. Conclusions and Perspectives

7 Molecular Systematics of Entomopathogenic Fungi

7.1. Introduction

7.2. Molecular Phylogenies of Fungi and the Origins of Entomopathogens

7.3. Species Recognition

7.4. Species-level Phylogenies of Entomopathogenic Fungi

7.5. Conclusions

8 Phylogenetics and Population Genetics of Entomopathogenic and Insect-parasitic Nematodes

8.1. Introduction

8.2. Phylogenetics

8.3. Population Genetics

8.4. DNA Bar Coding

8.5. DNA Markers Considered for Phylogenetic and Population Genetics Studies

8.6. Methodology

8.7. Co-phylogenesis and Cospeciation

8.8. Population Genetics Methods

PART III: HOST–PATHOGEN INTERACTIONS

9 Host–Virus Interactions

9.1. Introduction

9.2. Use of Viruses as Insect Pest Control Agents

9.3. Development of Baculoviruses for Foreign Gene Expression

9.4. Insect Defences Against Viruses

9.5. Baculovirus Pathogenesis

9.6. Baculovirus Host Range

9.7. Unclassified DNA Viruses

9.8. Mechanisms of Insect Virus Persistence

9.9. Conclusions and Future Prospects

10 Insect–Protozoa–Bacteria Associations: a Model System for Investigating Host–Parasite Interactions

10.1. Introduction

10.2. Trypanosomatid Protozoa and Tsetse Flies

10.3. Molecular Approaches and Their Application to Study Insect Host Immune Responses

10.4. Tsetse Endosymbionts

10.5. Control of Insect-borne Diseases

10.6. Current and Future Work

10.7. Conclusions

11 Methods in Investigating Nematode–Bacterium–Insect Symbiosis

11.1. Introduction

11.2. Molecular Tools to Study Entomopathogenic Nematodes

11.3. Basic Molecular Tools for the Study of Entomopathogenic Bacteria

11.4. Techniques to Investigate Bacteria–Nematode Mutualism

11.5. Techniques in Studying EPB Virulence

11.6. Conclusions and Future Prospects

PART IV: GENOMICS AND GENETIC ENGINEERING

12 Genetic Engineering of Bacteria to Improve Efficacy Using the Insecticidal Proteins of Bacillus Species

12.1. Introduction

12.2. Basic Biology of Bacillus thuringiensis

12.3. Insecticidal Proteins of Bacillus thuringiensis

12.4. Genetic Factors Regulating Insecticidal Proteins

12.5. Construction of Recombinant Bacteria

12.6. Conclusions and Future Prospects

13 Genomic Analysis of the Symbiotic and Entomopathogenic Photorhabdus Bacteria

13.1. Introduction

13.2. Sequencing and Annotation of Photorhabdus Genomes

13.3. Main Features of the P. luminescens Genome

13.4. Analogical Post-genomic Analysis

13.5. Post-genomic Analysis by a ‘Blind’ Approach

13.6. Conclusions and Future Perspectives

14 Genomics of Entomopathogenic Viruses

14.1. Introduction

14.2. General Concepts

14.3. Analyses of DNA and Protein Sequences

14.4. Genetic Modification of Baculoviruses

14.5. Conclusions

15 Genomics and Genetic Improvement of Entomopathogenic Nematodes

15.1. Introduction

15.2. Genomic Sequencing and Bioinformatics

15.3. Functional Genomics: Towards Deciphering of Genomics and ESTs Sequences

15.4. Genetic Improvement

15.5. Conclusion and Future Prospects

16 Entomopathonic Fungi and the Genomics Era

16.1. Introduction

16.2. Procedures for Isolating Pathogen DNA

16.3. Procedures for Isolating Pathogen RNA

16.4. Pulsed Field Gel Electrophoresis

16.5. Construction of Cloning and Expression Vector Components and Markers

16.6. Transformation Systems

16.7. Gene Cloning Strategies

16.8. Analysing Differential Gene Expression

16.9. EST Screening

16.10. Microarray Analysis

16.11. Targeted Gene Mutagenesis

16.12. RNA Interference

16.13. Conclusions

Index

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